LOCUS       sequence-237174-        4824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP R4-R3 entry plasmid, contains osTir1 for 3- or 4-component 
            MultiSite Gateway Pro assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4824)
  AUTHORS   Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, 
            Newgard CB
  TITLE     Creation of versatile cloning platforms for transgene expression and
            dCas9-based epigenome editing.
  JOURNAL   Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 
            10.1093/nar/gky1286.
  PUBMED    30590691
REFERENCE   2  (bases 1 to 4824)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286."
COMMENT     Sequence Label: KJ101: pMVP (R4-R3) osTir1
FEATURES             Location/Qualifiers
     source          1..4824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..726)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             777..2498
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="TYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFVG
                     NCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEEL
                     RMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEVE
                     DRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTLA
                     KILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYPL
                     CAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELRV
                     FPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRLC
                     ILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSIA
                     FAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKGC
                     QVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     CDS             2526..2555
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2562..2591
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2598..2627
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2646..2675
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2682..2711
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2718..2747
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2766..2795
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2802..2831
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2838..2867
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    2887..3010
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3065..3084)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3066..3084)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3089..3105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3089..3105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3207..3226)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3218..4027
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3285..3304)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4174..4762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4663..4682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgccat ggcattacaa aaaaaattag aagaattaga attagatgaa ggcggcacgt
      781 acttcccgga ggaggtggtg gagcacatct tcagcttcct gccggcgcag cgcgaccgca
      841 acacggtctc gctcgtctgc aaggtgtggt acgagatcga gaggctgagc cgccgcggcg
      901 tcttcgtggg caactgctac gccgtgcgcg ccggccgcgt cgccgcgcgg ttccccaacg
      961 tgcgggcgct cacggtgaag gggaagcccc acttcgccga cttcaacctc gtgccccccg
     1021 actggggcgg ctacgcgggg ccgtggatcg aggcggccgc gaggggatgc cacggcctgg
     1081 aggagctcag gatgaagcgg atggtggtgt ccgacgagag cctcgagctg ctggctcgct
     1141 cgttcccgcg gttcagggct cttgttctta tcagctgcga ggggttcagc actgacgggc
     1201 tagccgccgt cgcgagccat tgcaagcttc tgagggagtt ggatttgcag gaaaatgaag
     1261 tggaggatcg agggcctagg tggctttcct gcttccctga ttcctgcaca tcacttgtct
     1321 cattgaattt tgcctgcatc aaaggggagg ttaatgctgg ttcactggag agacttgtta
     1381 gcaggtcccc aaacctgcgg agtttgaggc tgaatcgatc tgtatcggta gatacacttg
     1441 caaagatact actgcgtacc cctaacttgg aggatttggg gacagggaat ttgacagatg
     1501 acttccaaac tgagtcctac tttaagctta ccagtgctct ggagaaatgc aagatgttga
     1561 ggagtttgtc tggattctgg gatgcttctc ctgtttgcct gtcatttatc taccccctgt
     1621 gtgctcaact gacaggattg aacttgagct atgcacccac acttgatgct tctgacctta
     1681 caaaaatgat tagccgctgt gtgaagctcc aacgcctttg ggtactggat tgtatctcgg
     1741 acaaaggctt gcaagtggtg gcctccagtt gcaaagactt gcaagaactc agggtatttc
     1801 catcagattt ctacgtagct ggttattctg cagtgacaga ggagggactt gttgcagtat
     1861 ccttgggctg tccaaaactg aactcactac tgtacttctg tcaccaaatg actaatgctg
     1921 cactagttac tgtcgccaag aactgtccaa atttcacacg attcagactt tgtattcttg
     1981 agccagggaa gcctgatgtt gtgacaagcc aaccattaga tgaaggcttt ggagctattg
     2041 ttcgtgagtg caagggatta caacgtttgt caatatctgg tcttctcaca gacaaagttt
     2101 tcatgtatat cgggaaatat gcaaaacaac ttgagatgct ttctatagca tttgctggtg
     2161 acagtgataa gggtatgatg catgttatga atggatgcaa gaatttaagg aaactggaga
     2221 taagagatag cccgtttggt gatgctgcac tcttggggaa ttttgctagg tacgagacaa
     2281 tgcgatccct ttggatgtca tcttgcaatg tcacgttaaa ggggtgccaa gtccttgcgt
     2341 caaagatgcc gatgctcaat gttgaggtca taaatgagcg ggatggtagc aatgaaatgg
     2401 aggaaaacca tggagatctg cctaaagtgg agaaattata tgtgtaccgc acaactgctg
     2461 gggcgaggga tgatgcacca aattttgtta aaatcctagt cgactccggt tctgctgcta
     2521 gtggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag ctaatctccg
     2581 aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac ggatcctcta
     2641 gaggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag ctaatctccg
     2701 aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac ggatcctcta
     2761 gaggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag ctaatctccg
     2821 aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac ggatccacta
     2881 gctaaacaac tttgtataat aaagttgaac gagaaacgta aaatgatata aatatcaata
     2941 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     3001 agtcactatg aatcaactac ttagatggta ttagtgacct gtactgcagg gcggccgcga
     3061 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3121 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     3181 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     3241 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3301 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     3361 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3421 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3481 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3541 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3601 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3661 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3721 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3781 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3841 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3901 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3961 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     4021 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     4081 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     4141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4321 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4741 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4801 ttgctggcct tttgctcaca tgtt
//
